Supplementary Figure 1: Restoration of P2 microbiota patterns of cesarean–delivered offspring via the maternal vaginal inoculant.
(a) Taxonomic classifications of microbiota colonizing the PN2 colon of C-section offspring that did not receive a maternal vaginal inoculant (noninoculated), C-section offspring that received a maternal vaginal inoculant (inoculated), and vaginally delivered offspring demonstrating greater similarity in microbiota community composition between inoculated and vaginally delivered offspring. N = 8 Caesarean delivered noninoculated mice; N = 24 Caesarean delivered inoculated mice; N = 29 Vaginally delivered mice. (b) Communities clustered using PCoA of the unweighted UniFrac distance matrix where each point corresponds to a sample collected from PN2 colons of vaginally delivered CTL and prenatal stress-exposed offspring, C-section noninoculated offspring, and C→C, C→S, S→C and S→S offspring. The percentage of variation explained by the PC is indicated on the axes. PCoA, principal coordinates analysis. N = 9 vaginally delivered control males; N = 20 vaginally delivered prenatal-stress exposed males; N = 8 noninoculated C-section delivered males; N = 6 C→C males; N = 9 C→S males; N = 6 S→C males; N = 5 S→S males. (c) Barplot of Unweighted UniFrac Distances showing significant differences in the community structure of noninoculated C-section offspring, vaginally delivered CTL and prenatal stress offspring, C→C, C→S, S→C, and S→S offspring at PN2 (One-way ANOVA, treatment, F6,386 = 9.253, P = 0.00000000175). N = 9 vaginally delivered control males; N = 20 vaginally delivered prenatal-stress exposed males; N = 8 noninoculated C-section delivered males; N = 6 C→C males; N = 9 C→S males; N = 6 S→C males; N = 5 S→S males. Data represented as mean ± SEM with individual UniFrac distance correlations overlaid. (d) Barplot (mean ± SEM) of Unweighted UniFrac Distances showing no significant difference in community structure of C→C, C→S, S→C, and S→S offspring at PN2. N = 6 C→C males; N = 9 C→S males; N = 6 S→C males; N = 5 S→S males. Data represented as mean ± SEM with individual UniFrac distance correlations overlaid. (e) Barplot (mean ± SEM) of the Shannon Diversity Index showing no effect of prenatal stress on community diversity in vaginally delivered offspring. Bacterial diversity is decreased in Noninoculated (NIC) offspring relative to vaginally delivered offspring (Vaginally delivered control vs. NIC, t15 = 2.774, P = 0.0142; Vaginally delivered prenatal stress exposed vs. NIC, t26 = 2.49, P = 0.0195, Unpaired t-Test). No difference in community diversity between vaginally delivered offspring and C-section offspring that received a maternal vaginal inoculant (C→C, C→S, S→C, and S→S offspring) (One-way ANOVA, main effect of treatment, F5,47 = 0.3447, P = 0.8831). N = 9 vaginally delivered control males; N = 20 vaginally delivered prenatal-stress exposed males; N = 8 noninoculated C-section delivered males; N = 6 C→C males; N = 9 C→S males; N = 6 S→C males; N = 5 S→S males. * P < 0.05. Data represented as mean ± SEM with individual data points overlaid.
